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Response of the microbial community to copper oxychloride in acidic sandy loam soil.

AIMS: Determining the response of different microbial parameters to copper oxychloride in acidic sandy loam soil samples using cultivation-dependent and direct microscopic techniques. METHODS AND RESULTS: Culturable microbial populations were monitored for 245 days in a series of soil microcosms spiked with different copper oxychloride concentrations. Microbial populations responded differently to additional Cu. Protistan numbers and soil metabolic potential decreased. Experiments with more soil samples revealed that metabolic potential was not significantly affected by < or =100 mg kg(-1) additional Cu. However, a negative impact on protista was noted in soil containing only 15 mg kg(-1) EDTA-extractable Cu. The negative impact on protistan numbers was less severe in soils with a higher phosphorous and zinc content. CONCLUSIONS: Bacterial populations responded differently, and protista were most sensitive to elevated Cu levels. Protistan numbers in soil from uncultivated land were higher and seemed to be more sensitive to additional Cu than the numbers of these organisms in soil originating from cultivated land. SIGNIFICANCE AND IMPACT OF THE STUDY: Protistan sensitivity to small increases in Cu levels demonstrates the vulnerability of the soil ecosystem to Cu perturbations, especially when the importance of protista as link in the flow of energy between trophic levels is considered.

Animals↗

Infancy.

Research on the psychology of infancy in the 1980s was prolific and diverse. At least four different perspectives have been taken. The first views the infant as information processor, adopts the methods and assumptions of experimental psychology, and focuses on such fundamental processes as attention, perception, emotion, learning and memory, search and exploration, and cognition. The second views the infant as an infant, as part of the natural world, and focuses on the important developmental milestones that uniquely characterize the period of infancy, such as walking, talking, forming relationships, and developing a sense of self. The third perspective views the infant as an eventual adult, emphasizes differences among individuals, and focuses on continuities across age and experiences such as biological risk factors, characteristics of rearing environments, and planned intervention that influence the kinds of adults infants will turn out to be. The fourth perspective focuses not on the infant per se, but on the general process of development; the issues examined concern the description and explanation of development, in terms of the existence of shifts and stages, the relationship of early attainments to later ones, the bidirectional influence of genes and environments, the timing of experience, individuals' contributions to their own experiences, and the reversibility of development. Synthesis across perspectives is difficult, but remains possible. The field of infancy research is a microcosm of psychology, but one that retains a view of the organism as holistic being. As such, the study of infancy produces insights of considerable relevance to psychology as a whole.

Attention↗

Effect of ethanol and methyl-tert-butyl ether on monoaromatic hydrocarbon biodegradation: response variability for different aquifer materials under various electron-accepting conditions.

Aquifer microcosms were used to determine how ethanol and methyl-tert-butyl ether (MtBE) affect monoaromatic hydrocarbon degradation under different electron-accepting conditions commonly found in contaminated sites experiencing natural attenuation. Response variability was investigated by using aquifer material from four sites with different exposure history. The lag phase prior to benzene, toluene, ethylbenzene, and xylenes (BTEX) and ethanol degradation was typically shorter in microcosms with previously contaminated aquifer material, although previous exposure did not always result in high degradation activity. Toluene was degraded in all aquifer materials and generally under a broader range of electron-accepting conditions compared to benzene, which was degraded only under aerobic conditions. The MtBE was not degraded within 100 d under any condition, and it did not affect BTEX or ethanol degradation patterns. Ethanol was often degraded before BTEX compounds and had a variable effect on BTEX degradation as a function of electron-accepting conditions and aquifer material source. An occasional enhancement of toluene degradation by ethanol occurred in denitrifying microcosms with unlimited nitrate; this may be attributable to the fortuitous growth of toluene-degrading bacteria during ethanol degradation. Nevertheless, experiments with flow-through aquifer columns showed that this beneficial effect could be eclipsed by an ethanol-driven depletion of electron acceptors, which significantly inhibited BTEX degradation and is probably the most important mechanism by which ethanol could hinder BTEX natural attenuation. A decrease in natural attenuation could increase the likelihood that BTEX compounds reach a receptor as well as the potential duration of exposure.

Biodegradation, Environmental↗

Use of isotopic and molecular techniques to link toluene degradation in denitrifying aquifer microcosms to specific microbial populations.

Microcosms were inoculated with sediments from both a petroleum-hydrocarbon (PHC)-contaminated aquifer and from a nearby pristine aquifer and incubated under anoxic denitrifying conditions with [methyl-13C]toluene. These microcosms served as a laboratory model system to evaluate the combination of isotope (13C-labeling of polar-lipid-derived fatty acids) and molecular techniques (16S rRNA-targeting gene probes) to identify the toluene-metabolizing population. After total depletion of toluene, the following bacterial phospholipid fatty acids (PLFA) were 13C-enriched: 16:1omega7c, 16:1omega7t, 16:0, cy17:0, and 18:1omega7c. Pure culture experiments demonstrated that these compounds were also found in PLFA profiles of PHC-degrading Azoarcus spp. (beta-Proteobacteria) and related species. The origin of the CO2 evolved in the microcosms was determined by measurements of stable carbon isotope ratios. Toluene represented 11% of the total pool of mineralized substrates in the contaminated sediment and 54% in the pristine sediment. The microbial community in the microcosm incubations was characterized by using DAPI staining and whole-cell hybridization with specific fluorescently labeled 16S rRNA-targeted oligonucleotide probes. Results revealed that 6% of the DAPI-stained cells in the contaminated sediment and 32% in the pristine sediment were PHC-degrading Azoarcus spp. In biotic control microcosms (incubated under denitrifying conditions, no toluene added), Azoarcus spp. cells remained at less than 1% of the DAPI-stained cells. The results show that isotope analysis in combination with whole-cell hybridization is a promising approach to identify and to quantify denitrifying toluene degraders within microbial communities.

Bacteria↗

Effects of chlorpyrifos in freshwater model ecosystems: the influence of experimental conditions on ecotoxicological thresholds.

Three experiments were conducted to determine the impact of the insecticide chlorpyrifos (single applications of 0.01 to 10 microg AI litre(-1)) in plankton-dominated nutrient-rich microcosms. The microcosms (water volume approximately 14 litres) were established in the laboratory under temperature, light regimes and nutrient levels that simulated cool 'temperate' and warm 'Mediterranean' environmental conditions. The fate of chlorpyrifos in the water column was monitored and the effects on zooplankton, phytoplankton and community metabolism were followed for 4 or 5 weeks. The mean half-life (t1/2) of chlorpyrifos in the water of the test systems was 45 h under 'temperate' conditions and about 30 h under 'Mediterranean' environmental conditions. Microcrustaceans (cladocerans and copepod nauplii) were amongst the most sensitive organisms. All three experiments yielded community NOEC (no observed effect concentrations) of 0.1 microg AI litre(-1), similar to those derived from more complex outdoor studies. Above this threshold level, responses and effect chains, and time spans for recovery, differed between the experiments. For example, algal blooms as an indirect effect from the impact of exposure on grazing organisms were only observed under the 'Mediterranean' experimental conditions. The relatively simple indoor test system seems to be sufficient to provide estimates of safe threshold levels for the acute insecticidal effects of low-persistence compounds such as chlorpyrifos. The robustness of the community NOEC indicates that this threshold level is likely to be representative for many freshwater systems.

Chlorpyrifos↗

Competition between ammonia-oxidizing bacteria and benthic microalgae.

The abundance, activity, and diversity of ammonia-oxidizing bacteria (AOB) were studied in prepared microcosms with and without microphytobenthic activity. In the microcosm without alga activity, both AOB abundance, estimated by real-time PCR, and potential nitrification increased during the course of the experiment. AOB present in the oxic zone of these sediments were able to fully exploit their nitrification potential because NH(4)(+) did not limit growth. In contrast, AOB in the alga-colonized sediments reached less than 20% of their potential activity, suggesting starvation of cells. Starvation resulted in a decrease with time in the abundance of AOB as well as in nitrification potential. This decrease was correlated with an increase in alga biomass, suggesting competitive exclusion of AOB by microalgae. Induction of N limitation in the oxic zone of the alga-colonized sediments and O(2) limitation of the majority of AOB in darkness were major mechanisms by which microalgae suppressed the growth and survival of AOB. The competition pressure from the algae seemed to act on the entire population of AOB, as no differences were observed by denaturing gradient gel electrophoresis of amoA fragments during the course of the experiment. Enumeration of bacteria based on 16S rRNA gene copies and d-amino acids suggested that the algae also affected other bacterial groups negatively. Our data indicate that direct competitive interaction takes place between algae and AOB and that benthic algae are superior competitors because they have higher N uptake rates and grow faster than AOB.

Ammonia↗

Survival, metabolic activity and conjugative interactions of indigenous and introduced streptomycete strains in soil microcosms.

The growth and activity of introduced (S. lividans TK24 pIJ673 and S. lividans TK23) and indigenous (S. griseus CAG17) streptomycete strains in soil was studied, under controlled conditions. The effect of environmental parameters such as temperature, soil water content and nutrient availability on the growth and activity of these strains, was studied using a highly dynamic fed-batch soil microcosm system. Using this new system, repeated cycles of active streptomycete growth were achieved, allowing long-term investigation of metabolic activity, plasmid stability and conjugative plasmid transfer. In long-term experiments, respiration rates and enzyme activity patterns matched the pattern of germination/sporulation cycles of the inoculants. In situ hybridisation, using fluorescently labelled oligonucleotides, also proved the presence of metabolically active streptomycete mycelia in sterile soil. Plasmid stability under varying temperatures and selective pressure was studied using the above system. In both sterile and non sterile amended antibiotic containing soil, no intraspecific transfer of plasmid pIJ673 from S. lividans TK24 to S. griseus CAG17 was detected. The soil microcosm system used, though, permitted detection of intraspecific conjugative transfer of this plasmid from S. lividans TK24 to S. lividans TK23 in soil.

Conjugation, Genetic↗

The incorrect interpretation.

Incorrect interpretations are unavoidable given that the nature of psychoanalytic inference is inexact, that the interactional pressures which are brought to bear may be intense, and that the analyst is human. However, the potentially harmful iatrogenic effect of incorrect interpretations upon the patient must be acknowledged and analysed. The patients develops a transitional relationship to the analyst's interpretations which is disrupted by interpretive errors and replaced by transference reactions to a pathogenic object. The patient's response repeats, in microcosm, his past affective and defensive responses to similar environmental lapses or impingements. Only if the analyst is aware of the patient's experience of the actuality of his error and rectifies it can the integrity of the patient's self-experience be restored, his response analysed in the transference, and reconstructions formulated about the antecedents of such reactions in the patient's past.

Defense Mechanisms↗

Toxicity to freshwater organisms from oils and oil spill chemical treatments in laboratory microcosms.

Toxicity and temporal changes in toxicity of freshwater-marsh-microcosms containing South Louisiana Crude (SLC) or diesel fuel and treated with a cleaner or dispersant, were investigated using Chironomus tentans, Daphnia pulex, and Oryzias latipes. Bioassays used microcosm water (for D. pulex and O. latipes) or soil slurry (for C. tentans) taken 1,7, 31, and 186 days after treatment. SLC was less toxic than diesel, chemical additives enhanced oil toxicity, the dispersant was more toxic than the cleaner, and toxicities were greatly reduced by day 186. Toxicities were higher in the bioassay with the benthic species than in those with the two water-column species. A separate experiment showed that C. tentans' sensitivity was intermediate to that of Tubifex tubifex and Hyallela azteca. Freshwater organisms, especially benthic invertebrates, thus appear seriously effected by oil under the worst-case-scenario of our microcosms. Moreover, the cleaner and dispersant tested were poor response options under those conditions.

Analysis of Variance↗

Microcosm studies for neutralization of hypolimnic acid mine pit lake water (pH 2.6).

Ten microcosms of 0.088 m3 water volume (0.3 m i.d. and 1.20 m height) were designed for neutralization studies representing hypolimnic ecosystem models for acid mine pit lakes. Sediment and water were collected from an acid lignite mine pit lake (Brandenburg, Germany) and filled into the microcosms. To determine the efficacy of controlled in situ organic carbon amendments as a possible neutralization method, sediment and water were treated with ethanol and Carbokalk with and without wheat straw. The water chemistry was monitored for 1 yr. At start-up and end of the experiments, the sedimentwas characterized. Iron and sulfate were removed with varying intensity from the water phase as a result of microbial iron and sulfate reduction together with a subsequent precipitation of unsoluble sulfide minerals to the sediment. The pH rose, and alkalinity generation and bacterial growth were observed. Neutralization rates were calculated using equivalents of accumulated total reduced inorganic sulfur together with the nonsulfidic reactive ferrous iron in the sediment. In the treated microcosms, the neutralization rates were between 6 and 15 equiv m(-2) a(-1). Carbokalk was most effective in stimulating growth of sulfate-reducing bacteria and probably also served as inoculum. With Carbokalk together with wheat straw, the pH increased from 2.6 to around 6.5 within the whole microcosm. The critical revision of the results indicates that the application of Carbokalk (approximately 3.9 kg m(-2)) together with the application of wheat straw (approximately 9.3 kg m(-2)) is most suitable for further experiments in outdoor enclosures (mesocosms). For that case, the prediction of the water quality for a lake water column after multiple lake turnover events is presented based on batch reaction simulation using the geochemical model PHREEQC.

Carbon↗

Effects of the phenylurea herbicide diuron on natural riverine microbial communities in an experimental study.

The effects of the phenylurea herbicide diuron (10 microgl(-1)) on natural riverine microbial communities were investigated using a three-week laboratory microcosm study. During the first six days, a latency period was observed both in the algal and the bacterial communities despite favorable abiotic conditions and independently of diuron exposure. From the second week, an intense algal bloom (chlorophyll a concentrations and cell abundances) was observed in the uncontaminated microcosms but not in the treated microcosms. The bloom stimulated the bacterial community and led to an increase in heterotrophic bacterial production ([3H]thymidine incorporation), activity (CTC reduction) and cell abundance. In parallel, shifts in bacterial community composition were recorded by polymerase chain reaction (PCR)-temporal temperature gradient gel electrophoresis (TTGE) analysis, whereas no major variation was detected using the fluorescent in situ hybridization (FISH) method. In the treated microcosms, the diuron acted not by damaging the initial communities but by inhibiting the algal bloom and indirectly maintaining constant bacterial conditions throughout the experiment. These inhibitory effects, which were recorded in terms of abundance, activity and diversity, suggest that exposure to diuron can decrease the recovery capacities of microbial communities and delay the resumption of an efficient microbial food web despite favorable environmental conditions.

Chlorophyll↗

Is interspecies hydrogen transfer needed for toluene degradation under sulfate-reducing conditions?

Sediments from a hydrocarbon-contaminated aquifer, where periodic shifts between sulfate reduction and methanogenesis occurred, were examined to determine whether the degradation of toluene under sulfate-reducing conditions depended on interspecies hydrogen transfer. Toluene degradation under sulfate-reducing conditions was inhibited by the addition of 5 mM sodium molybdate, but the activity was not restored upon the addition of an actively growing, hydrogen-using methanogen. Toluene degradation was not inhibited in microcosms where hydrogen levels were maintained at a level theoretically sufficient to inhibit toluene degradation if the process proceeded via interspecies hydrogen transfer. Finally, the addition of carbon monoxide, a potent inhibitor of hydrogenase activity, inhibited hydrogen but not toluene consumption in sulfate-reducing microcosms. These results suggest that toluene is degraded directly by sulfate-reducing bacteria without the involvement of interspecies hydrogen transfer. The sequence of experiments used to reach this conclusion could be applied to determine the role of interspecies hydrogen transfer in the degradation of a variety of compounds in different environments or under different terminal electron-accepting conditions.

Journal Article↗

Metabolic activity of bacterial cells enumerated by direct viable count.

The direct viable count (DVC) method was modified by incorporating radiolabeled substrates in microautoradiographic analyses to assess bacterial survival in controlled laboratory microcosms. The DVC method, which permits enumeration of culturable and nonculturable cells, discriminates those cells that are responsive to added nutrients but in which division is inhibited by the addition of nalidixic acid. The resulting elongated cells represent all viable cells; this includes those that are culturable on routine media and those that are not. Escherichia coli and Salmonella enteritidis were employed in the microcosm studies, and radiolabeled substrates included [methyl-3H]thymidine or [U-14C]glutamic acid. Samples taken at selected intervals during the survival experiments were examined by epifluorescence microscopy to enumerate cells by the DVC and acridine orange direct count methods, as well as by culture methods. Good correlation was obtained for cell-associated metabolic activity, measured by microautoradiography and substrate responsiveness (by the DVC method) at various stages of survival. Of the cells responsive to nutrients by the DVC method, ca. 90% were metabolically active by the microautoradiographic method. No significant difference was observed between DVC enumerations with or without added radiolabeled substrate.

Acridine Orange↗

Characterizing kinetics of transport and transformation of selenium in water-sediment microcosm free from selenium contamination using a simple mathematical model.

This study developed a seven-compartment model for predicting the fate of selenium (Se) in an aquatic environment containing a water-sediment boundary. Speciation of Se in water-sediment microcosms under microaerobic conditions was measured to evaluate first-order kinetics of Se transportation and transformation. The microcosm consisted of a 10-ml solution containing 1mM soluble Se as selenate (Se6+) or selenite (Se4+) and 8 g wet sediment that was free from Se contamination, sampled from the Senri, Yamato, or Yodo Rivers in Osaka, Japan. Stepwise reaction coefficients describing transportation and transformation were determined using an inverse method on this model which includes: selenate (Se(W)6+) and selenite (Se(W)4+) in ponded water; selenate (Se(S)6+) and selenite (Se(S)4+), elemental Se (Se0), organic Se (Se2-) in sediment; and gaseous Se (DMSe). During this 1-month experiment, soluble Se was transported from ponded water to the sediment and Se was transformed sequentially to other Se species through biochemical reactions. Experimental and kinetic analyses indicated quantitatively that the Yamato River microcosm, with its high organic matter content, had a high adsorption rate of soluble Se. The Yodo River microcosm had a low adsorption rate for Se6+ and a low Se reduction rate. The Senri River microcosm had an apparent high volatilization rate of DMSe. The model developed in this study is extremely useful for predicting fate of Se in aquatic environment in the field.

Biological Transport↗

[Interspecific interaction between Moina mongolica and Brachionus plicatilis].

In a laboratory experiment, Moina mongolica and Brachionus plicatilis were polycultured at four relatively inoculating densities, i.e., 0.06/0.30, 0.10/0.30, 0.30/0.30 and 0.60/0.30 (ind x ml(-1)/ind x ml(-1)), while M. mongolica monocultured at 0.06, 0.10, 0.30 and 0.60 ind x ml(-1) and B. plicatilis at 0.30 ind x ml(-1) were used as the control. Interspecific interference did exist between M. mongolica and B. plicatilis when these two species coexisted in a microcosm. In the polycultured microcosms, depressed population density of M. mongolica resulted from the coexistence of B. plicatilis, and M. mongolica maintained at extremely low density. In opposite, M. mongolica had negligibly negative influence on the population of B. plicatilis. Further experiment showed that the mortality of M. mongolica deprived of food for 120 h was 100%, while 90% of B. plicatilis survived after 144 h of food deprivation. The relatively strong capacity of B. plicatilis in tolerating starvation may be one of the important reasons for it wins the competition against M. mongolica.

Animals↗

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) by native microflora and combinations of white-rot fungi in a coal-tar contaminated soil.

Four white-rot fungi (Phanerochaete chrysosporium IMI 232175, Pleurotus ostreatus from the University of Alberta Microfungus Collection IMI 341687, Coriolus versicolor IMI 210866 and Wye isolate #7) and all possible combinations of two or more of these fungi, were incubated in microcosms containing wheat straw and non-sterile coal-tar contaminated soil to determine their potential to degrade polycyclic aromatic hydrocarbons (PAHs). Biotic and abiotic controls were prepared similarly and PAH concentrations remaining in each microcosm were determined after 8, 16 and 32 weeks by GC-MS following extraction with dichloromethane. The greatest PAH losses were in the biotic control, compared to small or negligible differences in microcosms inoculated with one or more fungi. These results suggest that in the biotic control native microorganisms colonised the straw added as organic substrate and degraded PAH as an indirect consequence of their metabolism. By contrast, in other microcosms, colonisation of straw by the natural microflora was inhibited because the straw was previously inoculated with fungi. Soil cultures prepared at the end of the experiment showed that though introduced fungi were still alive, they were unable to thrive and degrade PAH in such a highly contaminated soil and remained in a metabolically inactive form.

Bioreactors↗

Experimental tests of nutrient limitation in freshwater picoplankton.

On the basis of correlative studies, picoplankton in Calder Lake, New York, are apparently unaffected by seasonal fluxes in nutrient (N and P) levels. In this small eutrophic lake, picoplankton (<2.0- to 0.2-mum size) and nanoplankton (<20 to >2 mum) predominate. Microplankton (>20 mum) are typically least important. Experiments were conducted in situ to test whether N, P or N/P ratios affect the predominance of these smaller organisms. Manipulations were run in 4-liter microcosms during June, July, and August 1988, corresponding to periods of increasing stratification and nutrient depletion. Following nutrient additions, phytoplankton were harvested and fractionated into three size classes. Microplankton and nanoplankton were significantly stimulated by both N (2.5 to 50 muM) and P (1 to 20 muM) additions. The severity of nutrient limitation was greatest during July. Picoplankton responded less strongly to N additions and were never P limited. These field data support laboratory studies which indicate that bacterium-sized phytoplankton use nutrients more efficiently and are superior competitors within mixed communities.

Journal Article↗

Adaptation limits diversification of experimental bacterial populations.

Adaptation to a specific niche theoretically constrains a population's ability to subsequently diversify into other niches. We tested this theory using the bacterium Pseudomonas fluorescens, which diversifies into niche specialists when propagated in laboratory microcosms. Numerically dominant genotypes were allowed to diversify in isolation. As predicted, populations increased in fitness through time but showed a greatly decreased ability to diversify. Subsequent experiments demonstrated that niche generalists and reductions in intrinsic evolvability were not responsible for our data. These results show that niche specialization may come with a cost of reduced potential to diversify.

Adaptation, Physiological↗